Methods of identifying inhibitors of polypeptides-of-interest

Abstract

Methods of identifying inhibitors of polypeptide-of-interests are provided. Accordingly there is provided a method comprising: (a) expressing the polypeptide-of-interest in cells being xenogeneic to the polypeptide, wherein the polypeptide-of-interest is selected causing growth retardation of the cells when expressed therein; (b) contacting the cells expressing the polypeptide-of-interest with a test agent; (c) measuring growth of the cells following or concomitant with step (b), wherein a relief in the growth retardation is indicative that the test agent is an inhibitor of the polypeptide-of-interest; alternatively or additionally, the method is performed by: (a) expressing the polypeptide-of-interest in cells being xenogeneic to the polypeptide, wherein the polypeptide-of-interest is selected causing growth retardation of the cells when expressed therein; (b) culturing the cells expressing the polypeptide-of-interest under conditions which relieve the growth retardation, wherein the relief of the growth retardation is indicative of conditions that inhibit the polypeptide-of-interest.

1 . A method of identifying an inhibitor of a polypeptide-of-interest, the method comprising:
 (a) expressing the polypeptide-of-interest in cells being xenogeneic to the polypeptide, wherein said polypeptide-of-interest is selected causing growth retardation of said cells when expressed therein;   (b) contacting said cells expressing said polypeptide-of-interest with a test agent;   (c) measuring growth of said cells following or concomitant with step (b), wherein a relief in said growth retardation is indicative that said test agent is an inhibitor of said polypeptide-of-interest.   
     
     
         2 . The method of  claim 1 , further comprising synthesizing the test agent being the inhibitor of said polypeptide-of-interest. 
     
     
         3 . A method of identifying an inhibitor of a polypeptide-of-interest, the method comprising:
 (a) expressing the polypeptide-of-interest in cells being xenogeneic to the polypeptide, wherein said polypeptide-of-interest is selected causing growth retardation of said cells when expressed therein;   (b) culturing said cells expressing said polypeptide-of-interest under conditions which relieve said growth retardation, wherein said relief of said growth retardation is indicative of conditions that inhibit said polypeptide-of-interest.   
     
     
         4 . A method of identifying a polypeptide which is incompatible with cell growth or vitality, the method comprising:
 (a) expressing the polypeptide in cells being xenogeneic to the polypeptide; and   (b) measuring growth of said cells expressing the polypeptide compared to growth of control cells of the same origin not expressing the polypeptide under identical conditions, wherein when growth of said cells expressing the polypeptide is retarded compared to said control cells, the polypeptide is considered incompatible with cell growth or vitality.   
     
     
         5 . The method of  claim 3 , wherein said conditions comprise a molecule endogenously synthesized by said cells. 
     
     
         6 . The method of  claim 5 , further comprising synthesizing said molecule. 
     
     
         7 . The method of  claim 5 , further comprising contacting said cells expressing said polypeptide-of-interest with said molecule to identify said relief of said growth retardation, thereby validating said inhibitory activity of said molecule. 
     
     
         8 . The method of  claim 1 , wherein said polypeptide-of-interest is a recombinant peptide. 
     
     
         9 . The method of  claim 1 , wherein a selection of said polypeptide of-interest causing growth retardation of said cell culture is performed according to the method of  claim 1 . 
     
     
         10 . The method of  claim 1 , wherein said cells are microbial cells. 
     
     
         11 . The method of  claim 10 , wherein said microbial cells are bacterial cells. 
     
     
         12 . The method of  claim 11 , wherein said bacterial cells comprise Gram positive bacteria. 
     
     
         13 . The method of  claim 11 , wherein said bacterial cells comprise Gram negative bacteria. 
     
     
         14 . The method of  claim 10 , wherein said polypeptide is a human polypeptide. 
     
     
         15 . The method of  claim 1 , wherein expressing the polypeptide comprises induced expression. 
     
     
         16 . The method of  claim 1 , wherein said expressing is effected at least in part in a presence of a known inhibitor of said polypeptide so as to prevent death of said cells. 
     
     
         17 . The method of  claim 1 , wherein the polypeptide is not a chimeric polypeptide. 
     
     
         18 . The method of  claim 1 , wherein said polypeptide is a disease causing polypeptide. 
     
     
         19 . The method of  claim 1 , wherein said polypeptide-of-interest is selected from the group consisting of an ion channel and a protease. 
     
     
         20 . The method of  claim 1 , wherein said test agent comprises a nucleic acid sequence and wherein contacting refers to transforming said cells to express said nucleic acid sequence. 
     
     
         21 . The method of  claim 1 , being effected in high throughput configuration. 
     
     
         22 . The method of  claim 21 , wherein said test agent forms a part of a library. 
     
     
         23 . The method of  claim 18 , wherein said disease causing polypeptide comprises a viral polypeptide. 
     
     
         24 . The method of  claim 23 , wherein said viral polypeptide comprises an influenza polypeptide. 
     
     
         25 . The method of  claim 18 , wherein said influenza polypeptide is M2. 
     
     
         26 . An isolated bacterial cell expressing an influenza polypeptide.